摘要
目的观察原花青素对机械性创伤（MT）造成大鼠继发性心肌细胞及心脏功能损伤的影响，并探究其作用机制。方法将120只成年雄性SD大鼠按随机数字表法分为正常组、仓4伤组、创伤给药组（注射原花青素）和创伤溶剂组（注射等渗盐水），每组30只。制备大鼠MT模型。TUNEL染色后计算心肌细胞凋亡指数，经右颈总动脉向左心室内插管，记录左心室舒缩压力变化，通过流式细胞仪检测H9c2细胞注射原花青素前后活性氧自由基（ROS）数量变化，激光共聚焦显微镜下观察测定经Fluo-4AM标记的心肌细胞内ca“浓度变化，电子显微镜下观察心肌细胞超微结构改变。结果（1）创伤组、创伤给药组、创伤溶剂组MT后心肌细胞凋亡指数分别为（7．96±1．16）％、（2．44±0．70）％、（7．76±1．14）％，与正常组（0．64±0．59）％比较均明显增高（P〈0．01）；创伤溶剂组与创伤组比较差异无统计学意义，而创伤给药组与创伤组比较差异有统计学意义（P〈0．05）。（2）左心室内压上升最大速率（＋ap／dtmax）创伤组、创伤溶剂组分别为（3294．55±180．95）mmHg／s、（3337．80±220．11）mmHg／s，与正常组（4030．04±293．70）mmHg／s比较均明显下降（P〈0．01）；创伤给药组为（3926．72±364．95）mmHg／s，较创伤组升高（P〈0．05）。左心室内压下降最大速率（-dp／dtmax）创伤组、创伤溶剂组分别为（2294．55±298．69）mmHg／s、（2291．20±348．72）mmHg／s，与正常组的（2830．04±334．63）mmHg／s比较均明显下降（P〈0．05）；创伤给药组为（2801．52±266．25）mmHg／s，较创伤组升高（P〈0．05）。（3）与正常组比较，创伤组心肌组织可见染色质边集，颜色变深，肌间毛细血管上皮细胞可见明显空泡形成，原花青素干预后，肌间毛细血管上皮细胞核膜未见明显空泡形成；与创伤组比较，创伤给药组染色质边集，颜色变深的异常改变有所改善。结论原花青素能减轻MT后心肌细胞超微结构的病理性改变，抑制继发性心肌细胞凋亡，改善心脏功能，发挥心脏保护作用。
Objective To evaluate the effect of proanthocyanidins on cardiocyte apoptosis and car- diac dysfunction following mechanical trauma （MT） in rats and discuss its mechanism. Methods A total of 120 SD rats were allocated to normal group, trauma group, and trauma solvent group （ treated with pro- anthocyanidins） and trauma vehicle group （treated with saline solution） according to the random number table, with 30 rats per group. Small animal quantitative wound instrument was used to prepare the MT model in rats. Cardiocyte apoptosis was measured by TUNEL staining. Changes in left ventricular dias- tolic pressure were recorded by left ventricular intubation via the right common carotid artery. Reactive oxygen species （ROS） produced by H9c2 cells before and after proanthocyanidins administration was de- tected with flow cytometry. Level of Ca2＋ in cardiocytes marked by Fluo-4AM was determined with laser scanning confocal microscope. Ultrastructure changes in cardiocytes were observed under electronic micro- scope. Results After the induction of MT, apoptosis index of cardiocytes in trauma group, trauma solvent group and trauma vehicle group was （7.96 ± 1.16） % , （2. 44 ± 0. 70） % , and （7.76 ± 1.14） % respectively, far higher than （ 0. 64 ± 0. 59 ） % in normal group （ P 〈 0. 01 ）. By comparison with the trauma group, apoptosis index did not differ significantly in trauma vehicle group （P 〉 0. 05） but substan- tial reduction was detected in trauma solvent group （P 〈 0. 05）. The ＋ dp/dtmax of cardiac function in trau- ma and trauma vehicle groups was （3294. 55 ± 180. 95） mmHg/s and （3337.80 ±220. 11）mmHg/s respec- tively lowered from （4030. 04 ± 293.70） mmHg/s in normal group （ P 〈 0. 01 ）, and was （ 3926. 72 ± 364. 95 ） mrnHg/s in trauma solvent group lowered in contrast with trauma group （P 〈 0. 05 ）. The -dp/dtmx in trauma and trauma vehicle groups was （2294.55 ± 298. 69） mmHg/s and （ 2291.20 ± 348.72） mmHg/s respectively lowered significantly from （2830.04 ± 334.63 ） mmHg/s in normal group （P 〈 0. 05）, and was （2801.52 ± 266.25） mmHg/s increased significantly in contrast with trauma group （ P 〈 0. 05 ）. Compared with normal group, trauma group showed chromatin margination and condensation as well as marked vacu- olization in the epithelium from intramuscular capillaries. However, the pathological changes were im- proved after proanthocyanidins therapy. Conclusion Proanthocyanidins is effective in heart protection after the mechanical trauma by attenuating the pathological changes of myocardial ultrastructure and myo- cardial apoptosis and improving cardiac function.
出处
《中华创伤杂志》
CAS
CSCD
北大核心
2015年第4期347-352,共6页
Chinese Journal of Trauma
基金
国家自然科学基金资助项目（30971065）
大连市科技计划资助项目（2012E12SF074）
辽宁省教育厅基金资助项目（2009A194）
关键词
原花青素类
心脏损伤
氧化性应激
Proanthoeyanidins
Heart injuries
Oxidative stress
